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Mycobiology
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Seven New Records of Penicillium Species
Belonging to Section Lanata-Divaricata in Korea
Monmi Pangging, Thuong T. T. Nguyen & Hyang Burm Lee
To cite this article: Monmi Pangging, Thuong T. T. Nguyen & Hyang Burm Lee (2021): Seven New
Records of Penicillium Species Belonging to Section Lanata-Divaricata in Korea, Mycobiology, DOI:
10.1080/12298093.2021.1952814
To link to this article: https://doi.org/10.1080/12298093.2021.1952814
© 2021 The Author(s). Published by Informa
UK Limited, trading as Taylor & Francis
Group on behalf of the Korean Society of
Mycology.
Published online: 02 Aug 2021.
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RESEARCH ARTICLE
Seven New Records of Penicillium Species Belonging to Section
Lanata-Divaricata in Korea
Monmi Pangging, Thuong T. T. Nguyen and Hyang Burm Lee
Environmental Microbiology Lab, Department of Agricultural Biological Chemistry, College of Agriculture and Life Sciences,
Chonnam National University, Gwangju, Korea
ABSTRACT
Penicillium species are known to be ubiquitous environmental saprophytes. In the survey of
diversity of genus Penicillium, seven new records of Penicillium species belonging to section
Lanata-Divaricata were isolated from freshwater and soil samples collected from different
locations in Korea. Based on morphological characteristics and multilocus phylogenetic ana-
lysis of the rDNA internal transcribed spacer region (ITS), b-tubulin (BenA), and calmodulin
(CaM) genes, the isolated strains were identified as P. annulatum,P. camponotum,P. echinu-
lonalgiovense,P. globosum,P. limosum,P. onobense, and P. yunnanense, respectively. This
study presents detailed phylogenetic analyses and morphological descriptions of these spe-
cies that contribute to section Lanata-Divaricata in Korea.
ARTICLE HISTORY
Received 22 March 2021
Revised 31 May 2021
Accepted 28 June 2021
KEYWORDS
Freshwater; soil; section
Lanata-Divaricata; morph-
ology; phylogeny
1. Introduction
The genus Penicillium established by Link et al. in
1809 [1], has a worldwide distribution, isolated from
diverse substrates, including air, soil, freshwater, as
endophytes, insect specimens, indoor environments,
and food products [2,3]. Using phylogenetic
approach, often supported by phenotypic, physio-
logic and/or extrolite data, members of this genus is
divided into two subgenera, 32 sections and 89 ser-
ies [2]. Members of this genus are economically
important as it produce antibiotics, enzymes,
organic acids, alcohols and pharmaceuticals [4].
Whereas, some of them cause food spoilage, pro-
duce mycotoxins, and cause human and animal dis-
eases [5]. Currently, the genus contains 483
accepted species [2]. In Korea, more than 100
Penicillium species have been reported [6–12].
The Penicillium section Lanata-Divaricata was
established by Thom et al. in 1930 [13] for species
with biverticillate conidiophores that usually contain
an elongation of the conidiophore’s main axis and
metulae that diverge from axis to form an asymmet-
rical verticil. Thus, resulting in conidiophores to be
interpreted as monoverticillate, although they are in
most cases divergently branched biverticillate coni-
diophores (also termed divaricate). This group of
species mainly isolated from soil, some found from
air, and protea repens infructescence [3,14–16]. This
section is species-rich, with 56 species are accepted
until 2016 [14–19]. But the list is rapidly increasing
with many new Penicillium species recently
described from all over the world and added to sec-
tion Lanata-Divaricata. Up to date, 76 species of
Penicillium sect. Lanata-Divaricata have been
accepted [2]. Recently, P.soli,P.melanosporum,P.
siccitolerans and P.michoacanense, were discovered
from phosphate solubilizing soil in China and soil
samples as xerophilic in Mexico and Spain [20,21].
To our knowledge, only 16 species of Penicillium
section Lanata-Divaricata have been reported in
Korea until now [6,10,11]. Thus, the aim of this study
was to isolate, identify and describe the previously
unrecorded seven isolates found in soil and freshwater
samples collected from different locations in Korea, P.
annulatum,P. camponotum,P. echinulonalgiovense,P.
globosum,P. limosum,P. onobense,andP. yunnanense
based on multi-loci phylogenetic analysis of ITS, BenA
and CaM, and morphological data.
2. Materials and methods
2.1. Sampling and isolation
Details of freshwater and soil samples collected from
different locations in South Korea are shown in
Table 1. Serial dilution plating method was used to
isolate fungal strains; 1 g of soil or 1 mL of freshwater
sample was added to 9 mL sterile distilled water.
Approximately, 10
1
,10
2
,10
3
,and10
4
dilutions
were plated onto potato dextrose agar (PDA)
(Difco
TM
, Sparks, MD, USA) and malt extract agar
CONTACT Hyang Burm Lee hblee@jnu.ac.kr
ß2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology.
This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/),
which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
MYCOBIOLOGY
https://doi.org/10.1080/12298093.2021.1952814
(MEA) (Difco
TM
) containing 50 ppm streptomycin
solution. Plates were incubated at 25 Cfor
7–10 days. Then, colonies were transferred to new
PDA plates and incubated for 7 days at 25C.
For stock storage, pure isolates were maintained in
20% glycerol at 80 C and in PDA slant tubes in
the Environmental Microbiology Laboratory
Fungarium, Chonnam National University, Gwangju,
Korea. The isolated strains were also deposited in the
Collection of National Institute of Biological
Resources (NIBR), Incheon, and Culture Collection
of the Nakdonggang National Institute of Biological
Resources (NNIBR), Sangju, Korea as shown in
Table 1.
2.2. DNA extraction and PCR sequencing
Total genomic DNA was extracted directly from the
mycelia using Solg TM Genomic DNA Prep Kit
(Solgent Co. Ltd., Daejeon, Korea). The ITS rDNA
region was amplified with primer pairs ITS1/ITS4 or
ITS5/LR5 [22], BenA gene with T10/Bt2b [23]orT1/
Bt2b [23,24], and CaM gene were amplified with CF1/
CF4 [25], respectively. The PCR cycling programs
used for amplification follows: initial denaturation at
94 C for 4 min, followed by denaturing at 94 Cfor
1 min, then annealing (in case of ITS rDNA and
BenA)at56
C for 30 s (in case of CaM, annealing
temperature at 55 C for 50 s), extension for 2 min at
72 C, and a final 10 min elongation step at 72 C
which was followed by cooling at 4 Cfor30cycles.
PCR products were visualized in 1% (w/v) agarose gel
electrophoresis. The PCR products were purified with
the Accuprep PCR Purification Kit (Bioneer Corp.,
Daejeon, Korea). PCR products were sequenced using
the same PCR primers on an ABI PRISM3730XL
Genetic Analyzer (Applied Biosystems, CA, USA) at
Macrogen (Daejeon, Korea).
2.3. Phylogenetic analysis
All sequence data used in this study were obtained
from GenBank. Sequences were aligned with
Clustal_X version 2.1 [26] and were edited manually
with Bioedit version 7.2.6.0 [27]. Maximum likeli-
hood (ML) phylogenies were assessed using MEGA
7 software [28] and the Kimura 2-parameter model.
The p-distance substitution model with 1,000 boot-
strap replications was used for the assessment of the
reliability of internal branches. Penicillium glabrum
CBS 125543 (T) was used as outgroup. The sequen-
ces of the isolates in this study were deposited in
the database under the accession numbers shown in
Table 2.
2.4. Morphological studies
For characterization, the respective strains were ino-
culated into three points, namely Czapek yeast auto-
lysate agar (CYA), Blakeslee’s malt extract agar
(MEA), and yeast extract sucrose agar (YES), and
incubated at 25 C for 7 days [29]. For morpho-
logical observations, fragments of mycelia were
removed from the cultures and placed on micro-
scope slides with lactic acid (60%). An Olympus
BX51 microscope (Olympus, Tokyo, Japan) was
used to capture digital images.
3. Results
3.1. Phylogenetic analysis
A BLASTn search on the ITS, BenA, and CaM
regions of the isolates were obtained. ITS regions of
CNUFC ULD4-13, CNUFC ULD4-3, CNUFC
MDW-11, CNUFC WG2-7, CNUFC DMS3-17,
CNUFC WG2-1, and CNUFC WNP1 showed simi-
larities of 99.8% (498/499 bp), 99.8% (485/486 bp),
99.8% (501/502 bp), 99.8% (488/489 bp), 100% (533/
533 bp), 100% (533/533 bp), and 99.8% (481/482 bp)
with P.annulatum (NR_138303), P.camponotum
(NR_158823), P.echinulonalgiovense (GU981587), P.
globosum (KY495018), P.limosum (NR_111496), P.
onobense (NR_111497), and P.yunnanense
(KY494989), respectively. In a BLASTn search with
BenA sequence, the isolate CNUFC ULD4-13,
Table 1. Information of Penicillium isolates used in this study.
Species Strain Culture Collection Source Location
Penicillium annulatum CNUFC ULD4-13 IMYKFGC000000043 Rhizosphere soil Seonginbong Peak, Ulleung Island, Korea
(3730004”N 13051023”E)
Penicillium camponotum CNUFC ULD4-3 QWJQFGC000000303 Rhizosphere soil Seonginbong Peak, Ulleung Island, Korea
(3730004”N 13051023”E)
Penicillium
echinulonalgiovense
CNUFC MDW-11 QWJQFGC000000469 Freshwater Mt. Mudeung, Wonhyo valley, Gwangju,
Korea (3508021”N 12659033”E)
Penicillium globosum CNUFC WG2-7 NNIBRFG25626 Freshwater Jeongdori Gugyedeung, Wando, Korea
(341901.20”N 1264500.00”E)
Penicillium limosum CNUFC DMS3-17 QWJQFGC000000008 Damp soil Sandeok-ri, Goseomyeon, Damyang, Korea
(3519016.1”N 12659017.401”E)
Penicillium onobense CNUFC WG2-1 NNIBRFG9318 Freshwater Jeongdori Gugyedeung, Wando, South
Korea (341901.20”N 1264500.00”E)
Penicillium yunnanense CNUFC WNP1 NNIBRFG25628 Freshwater Doam-gil, Wando, Korea (341901.20”N
1264500.00”E)
2 M. PANGGING ET AL.
Table 2. GenBank accession numbers for fungal strains used in this study.
Species Strain
GenBank Accession no.
ITS BenA CaM
P.alagoense URM 93058 (T) MK804503 MK802333 MK802336
P.annulatum CBS 135126 (T) JX091426 JX091514 JX141545
P.annulatum CNUFC ULD4-13 MZ234244 MZ284832 MZ284839
P.araracuarense CBS 113149 (T) GU981597 GU981642 KF296373
P.bissettii CBS 140972 (T) KT887845 KT887806 KT887767
P.brasilianum CBS 253.55 (T) GU981577 GU981629 MN969239
P.brefeldianum CBS 235.81 (T) AF033435 GU981623 AB667857
P.camponotum CBS 140982 (T) KT887855 KT887816 KT887777
P.camponotum CNUFC ULD4-3 MZ234245 MZ284833 MZ284840
P.caperatum CBS 443.75 (T) KC411761 GU981660 KF296392
P.cataractum CBS 140974 (T) KT887847 KT887808 KT887769
P.cluniae CBS 326.89 (T) KF296406 KF296471 KF296402
P.coeruleum CBS 141.45 (T) GU981606 GU981655 KF296393
P.cremeogriseum CBS 223.66 (T) GU981586 GU981624 KF296403
P.curticaule CBS 135127 (T) FJ231021 JX091526 JX141536
P.echinulonalgiovense CBS 328.59 (T) GU981587 GU981631 KX961269
P.echinulonalgiovense CNUFC MDW-11 MZ234246 MZ284834 MZ284841
P.ehrlichii CBS 324.88 (T) AF033432 GU981652 KF296395
P.elleniae CBS 118135 (T) GU981612 GU981663 MN969254
P.excelsum DTO 357-D7 (T) KR815341 KP691061 KR815342
P.fructuariae-cellae CBS 145110 (T) MK039434 KU554679 MK045337
P.glaucoroseum CBS 138908 (T) MN431390 MN969383 MN969257
P.globosum CBS 144639 (T) KY495014 KY495123 KY494954
P.globosum CNUFC WG2-7 MZ234247 MZ284835 MZ284842
P.griseoflavum CGMCC 3.18799 (T) KY495011 KY495120 KY494951
P.guangxiense CBS 144526 (T) KY494986 KY495095 MN969332
P.hainanense CBS 144527 (T) KY495009 KY495118 KY494949
P.infrabuccalum CBS 140983 (T) KT887856 KT887817 KT887778
P.janthinellum CBS 340.48 (T) GU981585 GU981625 MN969268
P.javanicum CBS 341.48 (T) GU981613 GU981657 GU981613
P.koreense KACC 47721 (T) KJ801939 KM000846 N/A
P.laevigatum CGMCC 3.18801 (T) KY495015 KY495124 KY494955
P.levitum CBS 345.48 (T) GU981607 GU981654 KF296394
P.limosum CBS 339.97 (T) GU981568 GU981621 KF296398
P.limosum CNUFC DMS3-17 MZ234248 MZ284836 MZ284843
P.lineolatum CBS 188.77 (T) GU981579 GU981620 MN969272
P.ludwigii CBS 417.68 (T) KF296409 KF296468 MN969273
P.malacosphaerulum CBS 135120 (T) FJ231026 JX091524 JX141542
P.mariae-crucis CBS 271.83 (T) GU981593 GU981630 KF296374
P.meloforme CBS 445.75 (T) KC411762 GU981656 KF296396
P.ochrochloron CBS 357.48 (T) GU981604 GU981672 KF296378
P.onobense CBS 174.81 (T) GU981575 GU981627 KF296371
P.onobense CNUFC WG2-1 MZ234249 MZ284837 MZ284844
P.ortum CBS 135669 (T) JX091427 JX091520 JX141551
P.panissanguineum CBS 140989 (T) KT887862 KT887823 KT887784
P.paraherquei CBS 338.59 (T) AF178511 KF296465 KF296372
P.pedernalense CBS 140770 (T) KU255398 KU255396 N/A
P.piscarium CBS 362.48 (T) GU981600 GU981668 KF296379
P.pulvillorum CBS 280.39 (T) AF178517 GU981670 KF296377
P.raperi CBS 281.58 (T) AF033433 GU981622 MN969291
P.reticulisporum NRRL 3447 (T) AF033437 GU981665 KF296391
P.rolfsii CBS 368.48 (T) JN617705 GU981667 KF296375
P.rubriannulatum CBS 144641 (T) KY495029 KY495138 KY494969
P.setosum CBS 144865 (T) KT852579 MF184995 MH105905
P.simplicissimum CBS 372.48 (T) GU981588 GU981632 KF296368
P.skrjabinii CBS 439.75 (T) GU981576 GU981626 KF296370
P.soliforme CBS 144482 (T) KY495038 KY495147 MN969337
P.spinuliferum CBS 144483 (T) KY495040 KY495149 MN969338
P.subrubescens DTO 188-D6 (T) KC346350 KC346327 KC346330
P.svalbardense CBS 122416 (T) GU981603 KC346325 KC346338
P.tanzanicum CBS 140968 (T) KT887841 KT887802 KT887763
P.terrarumae CBS 131811 (T) MN431397 KX650295 MN969323
P.vasconiae CBS 339.79 (T) GU981599 GU981653 KF296386
P.wotroi CBS 118171 (T) GU981591 GU981637 KF296369
P.yunnanense CBS 144485 (T) KY494990 KY495099 KY494930
P.yunnanense CNUFC WNP1 MZ234250 MZ284838 MZ284845
Bold letters indicate isolates and accession numbers determined in our study.
CBS: Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; CNUFC: Chonnam National University Fungal
Collection (Gwangju, South Korea); DTO: Working collection of the Applied and Industrial Mycology department
housed at the Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands; 7); KACC: Korean Agricultural Culture
Collection, National Institute of Agricultural Biotechnology, Rural Development Administration (Suwon, South Korea);
URM: Culture collection (WCDM 604) hosted at the Departmento de Micologia of the Universidade Federal de
Pernambuco, Recife, Brazil. T: ex-type strain; N/A: Not available.
MYCOBIOLOGY 3
CNUFC ULD4-3, CNUFC MDW-11, CNUFC
WG2-7, CNUFC DMS3-17, CNUFC WG2-1, and
CNUFC WNP1 showed similarity 99.6% (470/
472 bp), 99.17% (479/483 bp), 98.80% (411/416 bp),
100% (416/416 bp), 99.79% (466/467 bp), 100% (480/
480 bp), and 99.76% (411/412 bp) with sequences of
P.annulatum CV0037 (JX091514), P.camponotum
(KT887819), Penicillium sp. YD-2017a (KY495150),
P.globosum (KY495127), P.limosum (GU981621),
P.onobense (GU981627), and P.yunnanense
(KY495099), respectively. Similarly, CaM sequence
of CNUFC ULD4-13, CNUFC ULD4-3, CNUFC
MDW-11, CNUFC WG2-7, CNUFC DMS3-17,
CNUFC WG2-1, and CNUFC WNP1 showed simi-
larities of 98.53% (536/544 bp), 98.64% (436/442 bp),
98.51% (398/404 bp), 99.5% (402/404 bp), 100%
(407/407 bp), 99.75% (405/406 bp), and 99.75% (405/
406 bp) with P.annulatum (JX141547), P.campono-
tum (KT887781), Penicillium sp. YD-2017a
(KY494981), P.globosum (KY494958), P.limosum
(KF296398), P.onobense (KF296371), and P.yunna-
nense (KY494930), respectively. Phylogenetic tree
based on the combined sequence data of the three
loci, ITS, BenA, and CaM revealed that the seven
isolated strains were identical to P.annulatum,P.
camponotum, P.echinulonalgiovense,P.globosum,P.
onobense,P.limosum, and P.yunnanense (Figure 1).
3.2. Taxonomy
3.2.1. Taxonomy of CNUFC ULD4-13
Penicillium annulatum Visagie & K. Jacobs,
Mycological Progress 14 (10/96): 14 (2015) (Figure 2).
Figure 1. Phylogenetic tree of Penicillium annulatum CNUFC ULD4-13, P. camponotum CNUFC ULD4-3, P. echinulonalgiovense
CNUFC MDW-11, P. globosum CNUFC WG2-7, P. limosum CNUFC DMS3-17, P. onobense CNUFC WG2-1, and P. yunnanense
CNUFC WNP1 and related species belonging to section Lanata-Divaricata based on maximum likelihood analysis of the com-
bined ITS, BenA,andCaM sequences. Numbers at the nodes indicate the bootstrap values (>70%) from 1,000 replicates. The
bar indicates the number of substitutions per nucleotide. The study isolates are shown in bold and different colors.
4 M. PANGGING ET AL.
Colony characteristics: CYA 25 C, 7 days: colo-
nies were moderately deep, radially and concentric-
ally sulcate, with ring-like appearance because of
sporulating and nonsporulating areas, with low mar-
gins, light-green to white mycelia, floccose texture,
sparse to moderately dense sporulation, no soluble
pigment, reverse light to grayish orange, and
reached 40–46 mm in diameter. MEA 25 C, 7 days:
colonies were low to moderately deep, plane, low,
irregular margins, light-green to white mycelia, floc-
cose texture, moderately dense to dense sporulation,
no exudate, no soluble pigment, reverse dull-green
to greenish white, and reached 42–47 mm in diam-
eter. YES 25 C, 7 days: colonies were radially sul-
cate, with ring-like appearance because of
sporulating and nonsporulating areas but less than
those on CYA, low, narrow margins, white mycelia,
floccose texture, sparse to moderate sporulation, no
exudate and soluble pigment, reverse dull-yellow to
yellowish white, and reached 41–49 mm in diameter.
Micromorphology: Conidiophores were bi- and
terverticillate, 165–720 3–4.5 mm. Metulae were
appressed to divergent, 7.8–20 2.5–4.5 mm.
Phialides were ampulliform 3–5 per metula,
6.2–82–3.5 mm. Conidia were globose to subglo-
bose, 2.5–32–3mm.
3.2.2. Taxonomy of CNUFC ULD4-3
Penicillium camponotum Visagie, David Clark, &
Seifert, Persoonia 36: 271 (2016) (Figure 3).
Colony characteristics: CYA 25 C, 7 days: colo-
nies were moderately deep, lightly radially sulcate,
narrow, entire margins, white mycelia, floccose tex-
ture, sparse to absent sporulation, no soluble pig-
ments, reverse yellowish white to pale-yellow, and
reached 20–22 mm in diameter. MEA 25 C, 7 days:
Figure 2. Morphology of Penicillium annulatum. (A, D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–I) Conidiophores; (J) Conidia (scale bars: G–J¼20 lm).
MYCOBIOLOGY 5
colonies were plane, entire margins, white mycelia,
floccose texture; moderately dense sporulation, no
soluble pigments, no exudates, reverse pinkish gray,
and reached 21–26 mm in diameter. YES 25 C,
7 days: colonies were moderately deep, radially, and
concentrically sulcate, with low, wide, entire mar-
gins, white mycelia, floccose texture, moderately
dense sporulation, no soluble pigments, reverse
light-brown, and reached 21–23 mm in diameter.
Micromorphology: Conidiophores were biverticil-
late, 212–610 2.5–3.5 mm. Metulae were divergent,
12.2–25.6 2.5–4mm. Phialides were ampulliform,
3–6 per metula, 8.2–11.4 2.5–3.5 mm. Conidia were
globose, 2.2–3.5 2.1–3.4 mm.
3.2.3. Taxonomy of CNUFC MDW-11
Penicillium echinulonalgiovense S. Abe ex
Houbraken & R.N. Barbosa, Antonie van
Leeuwenhoek 111 (10): 1895 (2018) (Figure 4).
Colony characteristics: CYA 25 C, 7 days: colonies
were moderately deep and lightly radially sulcate, with
low, narrow, entire margins, white mycelia, floccose
texture, sparse to absent sporulation, no soluble pig-
ments, reverse yellowish white to pale-yellow, and
reached 18–22 mm in diameter. MEA 25 C, 7 days:
colonies were low to moderately deep and plane, with
low, irregular margins, light-green to white mycelia,
floccose texture, moderately dense to dense sporula-
tion, no exudate, no soluble pigment, reverse dull-
green to greenish white, and reached 20–23 mm in
diameter. YES 25 C, 7 days: colonies were low to
moderately deep, sunken at center, radially and con-
centrically sulcate, with low, narrow, entire margins,
whitemycelia,floccosetexture,sparsetomoderately
dense sporulation, no exudate and soluble pigment,
reverse dull-green, and reached 17–20 mm in diameter.
Micromorphology: Conidiophores were biverticil-
late, 56–210 2.2–3.0 mm. Metulae were divergent,
Figure 3. Morphology of Penicillium camponotum. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–I) Conidiophores; (J) Conidia (scale bars: G–J¼20 lm).
6 M. PANGGING ET AL.
10.7–18.8 2.2–3.1 mm. Phialides were ampulliform,
2–6 per metula, 7.1–10.9 2.5–3.0 mm. Conidia
were echinulate, globose to subglobose
2.5–3.9 2.3–3.7 mm.
3.2.4. Taxonomy of CNUFC WG2-7
Penicillium globosum L. Cai, Houbraken, & X.Z.
Jiang, Cladistics 35 (5): 529 (2018) (Figure 5).
Colony characteristics: CYA 25 C, 7 days: colo-
nies were sunken in center, radially and concentric-
ally sulcate, low, narrow, entire margins, blueish
white mycelia, floccose texture, sparse to moderately
dense sporulation, no soluble pigment, reverse pale-
orange to apricot, and reached 20–22 mm in diam-
eter. MEA 25 C, 7 days: colonies were low, irregular
margins, white mycelia, floccose texture, sparse to
moderately dense sporulation, no exudate, no sol-
uble pigment, reverse grayish green, and reached
20–24 mm in diameter. YES 25 C, 7 days: colonies
were deep, raised at center, radially and
concentrically sulcate, low, narrow, entire margins,
white mycelia, floccose, sulcate texture, sparse to
moderately dense sporulation, no exudate, no sol-
uble pigment, reverse grayish orange, and reached
16–19 mm in diameter.
Micromorphology: Conidiophores were mono-
and biverticillate, 42–162 3.6–4mm. Metulae were
appressed to divergent, 7.6–20.2 2.7–4mm.
Phialides were ampulliform, 3–9 per metula,
7.8–12.6 2.2–4mm. Conidia were globose to sub-
globose, 2.9–4.1 2.1–3.8 mm.
3.2.5. Taxonomy of CNUFC DMS3-17
Penicillium limosum S. Ueda, Mycoscience 36 (4):
451 (1995) (Figure 6).
Colony characteristics: CYA 25 C, 7 days: colo-
nies were radially sulcate at center, with light gray-
ish green to white floccose surface, no exudate and
soluble pigment, reverse brown to brownish orange,
and reached 39–44 mm in diameter. MEA 25 C,
Figure 4. Morphology of Penicillium echinulonalgiovense. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies
on Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–I) Conidiophores; (J) Conidia (scale bars: G–J¼20 lm).
MYCOBIOLOGY 7
7 days: colonies were plain, white to grayish green
mycelia, with sparse to moderately dense sporula-
tion, no soluble pigment, reverse grayish yellow, and
reached 35–41 mm in diameter. YES 25 C, 7 days:
colonies were deep, raised at center, radially and
concentrically sulcate, with low, narrow, entire mar-
gins, white mycelia, floccose, sulcate texture, sparse
to moderately dense sporulation, no exudate, no sol-
uble pigment, reverse grayish orange, and reached
36–40 mm in diameter.
Micromorphology: Conidiophores were biverticil-
late, 73–252 mm. Metulae were appressed to divergent,
14.8–20.2 3–4mm. Phialides were ampulliform, 5–6
per metula, 8–12.6 2.2–3mm. Conidia were globose
to subglobose, 2.7–3.2 2.4–3mm.
3.2.6. Taxonomy of CNUFC WG2-1
Penicillium onobense C. Ram
ırez & A.T. Mart
ınez,
Mycopathologia 74 (1): 44 (1981) (Figure 7).
Colony characteristics: CYA 25 C, 7 days: colo-
nies were velvety, radiately wrinkled, dirty white,
entire margins, white mycelia, velutinous texture,
moderately dense sporulation, no soluble pigments
and exudates, reverse pale-brownish orange, and
reached 19–22 mm in diameter. MEA 25 C, 7 days:
colonies were low, with low, irregular margins,
green mycelia, grayish green at center, floccose tex-
ture, sparse to moderately dense sporulation, no
exudate, no soluble pigment, reverse grayish green
with light orange circle at center, and reached
36–43 mm in diameter. YES 25 C, 7 days: colonies
were moderately deep, radially and concentrically
sulcate, with low, wide, entire margins, white myce-
lia, floccose texture, moderately dense sporulation,
no soluble pigments, no exudates, reverse light-
brown, and reached 18–23 mm in diameter.
Micromorphology: Conidiophores were biverticil-
late, 73–310 mm. Metulae were divergent,
Figure 5. Morphology of Penicillium globosum. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–I) Conidiophores; (J) Conidia (scale bars: G–J¼20 lm).
8 M. PANGGING ET AL.
10.2–13.1 3.3–5mm. Phialides were ampulliform,
3–6 per metula, 8.3–12.2 2.2–3mm. Conidia were
elliptical to subglobose, 2.3–4.1 2.1–4.1 mm.
3.2.7. Taxonomy of CNUFC WNP1-1
Penicillium yunnanense L. Cai & X.Z. Jiang,
Cladistics 35 (5): 545 (2018) (Figure 8).
Colony characteristics: CYA 25 C, 7 days: colo-
nies were moderately deep, raised at center, radially
sulcate, with low, narrow, entire margins, white
mycelia, velutinous texture, moderately dense sporu-
lation, no soluble pigments, no exudates, reverse
greenish gray to dull-green, and reached 22–24 mm
in diameter. MEA 25 C, 7 days: colonies were mod-
erately deep, radially sulcate, with low, narrow,
irregular margins, white mycelia, floccose texture,
sparse to moderately dense sporulation, no exudate,
no soluble pigment, reverse pale to yellowish gray,
pale yellowish circles at center, and reached
23–24 mm in diameter. YES 25 C, 7 days: colonies
were low to moderately deep, sunken at center, radi-
ally and concentrically sulcate, with low, narrow,
entire margins, white mycelia, floccose texture,
sparse to moderately dense sporulation, no exudate,
no soluble pigment, reverse dull-green, and reached
26–28 mm in diameter.
Micromorphology: Conidiophores were mono-
and biverticillate, 22–126 2–3mm. No metulae
were observed. Phialides were ampulliform,
6.6–14.1 2–4mm. Conidia were broadly ellipsoidal
to ellipsoidal, 3–42.5–3.5 mm.
4. Discussion
This study advanced our understanding of
Penicillium sect. Lanata-Divaricata and contributed
seven new records for Korea.
Figure 6. Morphology of Penicillium limosum. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–J) Conidiophores; (K) Conidia (scale bars: G–K¼20 lm).
MYCOBIOLOGY 9
Although ITS rDNA is designated as the univer-
sal primers available and most widely sequenced
marker for fungi [30], but not variable enough for
distinguishing all closely related species in
Penicillium [3,31]. Thus, additional secondary
marker such as BenA and CaM were proposed and
found to be useful for the accurate identification of
Penicillium species [2,3,15]. Phylogenetic tree based
on combined ITS-BenA-CaM sequences indicated
that seven strains belonging to the series Janthinella,
Rolfsiorum and Simplicissima in section Lanata-
Divaricata (Figure 1).
The isolates CNUFC ULD4-13 was well placed
within the clade of P. annulatum in series
Rolfsiorum (Figure 1). The isolate was morphologic-
ally similar to description of P.annulatum [15] with
respect to producing bi- and terverticillate conidio-
phores, ampuiliform phialides, and roughened
globose to subglobose conidia (Figure 2). Previous
studies showed P.annulatum to be isolated from air
sample, soil, and mite in Protea repens infructes-
cence and Stellenbosch in South Africa [15]. There
are no studies on extrolites produced by P.annula-
tum. Similarly, the phylogenetic placement and mor-
phological characteristics of isolates CNUFC ULD4-
3 into series Rolfsiorum (Figure 1), matched previ-
ously with described species, P.camponotum [16].
Penicillium camponotum isolated from carpenter
ants from New Brunswick, Canada, and ant nest in
Picea abies from Germany, was reported to produce
andrastin A, B, and C, citrinalin, mangrovamides,
marcfortine A and B, and patulin [16]. In this study,
P.camponotum was isolated from soil samples. The
isolate CNUFC MDW-11 was grouped with P.echi-
nulonalgiovense in series Simplicissima (Figure 1)
and shared morphology similar to the previous
Figure 7. Morphology of Penicillium onobense. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–I) Conidiophores; (J) Conidia (scale bars: G–J¼20 lm).
10 M. PANGGING ET AL.
descriptions. Penicillium echinulonalgiovense was
first isolated from soil samples in Japan without a
Latin diagnosis, later validated by Barbosa et al.
[32,33]. Penicillium echinulonalgiovense was also iso-
lated from bee pollen and nest of Melipona scutella-
ris located at Recife, Pernambuco in Brazil; soil
samples from Australia, China, Hong Kong,
Indonesia, USA, Madagascar, and Malaysia; and
industrial installations in Netherlands [33].
Penicillium echinulonalgiovense produces andrastin
A, xanthoepocin, and pulvilloric acid [2,33].
Similarly, CNUFC WG2-7 was clustered within the
same clade as P.globosum in series Simplicissima
(Figure 1), described by Diao et al. [34]. The isolate
CNUFC WG2-7 was morphologically similar to P.
globosum with only differences in the number of
phialides per metula. According to Diao et al. [34],
3–19 phialides per metula whereas the isolate
CNUFC WG2-7 consists of 3–9 phialides per
metula. P.globosum was isolated from an acidic soil
in China and Australia; rainforest soil from
Malaysia; industrial installations in Netherlands; and
soil in citrus grove in Florida, USA [34]. Also, the
morphological characteristics of CNUFC DMS3-17
were similar to the previous descriptions [35] and
was also clustered in P.limosum CBS 339.97 (type)
in series Janthinella (Figure 1). Earlier, Penicillium
limosum have been reported to be found only in
marine sediment in Nagasaki Prefecture, Japan [35].
In this study, we isolated P.limosum from damp
soil samples. Penicillium limosum was reported to
produce a sexual state [35] but no sexual state is
found in CNUFC DMS3-17 isolate. Based on previ-
ous descriptions, CNUFC WG2-1 shared similar
morphology and placed in series Simplicissima
(Figure 1). Penicillium onobense isolated from soil
and andosol in Navarra, Spain [36], produces brefel-
din A, janthitrems/shearinins, and 2-(4-
Figure 8. Morphology of Penicillium yunnanense. (A,D) Colonies on Czapek yeast autolysate agar (CYA); (B,E) Colonies on
Blakeslee’s malt extract agar (MEA); (C,F) Colonies on yeast malt extract agar (YES). (A–C: obverse view, D–F: reverse view).
(G–J) Conidiophores; (K) Conidia (scale bars: G–K¼20 lm).
MYCOBIOLOGY 11
hydroxyphenyl)-2-oxoacetaldehyde oxime [2]. The
isolate CNUFC WNP1 was placed in series
Janthinella (Figure 1) and also shared similar mor-
phological characters with previous descriptions
[34]. P.yunnanense was isolated from acidic soil in
China [34]. Interestingly, this study isolated P.echi-
nulonalgiovense,P.globosum,P.onobense, and P.
yunnanense from freshwater for the first time.
This study on the isolation and description of
seven new records of Penicillium sect. Lanata-
Divaricata from freshwater and soil samples adds to
our knowledge on fungal biodiversity. Different
environmental sources such as bees, ants, flowers,
leaves, nut kernels, and shells studies are needed in
Korea considering the increasing evidence for eco-
logical specialization in Penicillium species.
Additional studies on the production of extracellular
enzymes, antimicrobial compounds, and extrolites
are still needed in the genus Penicillium.
Disclosure statement
No potential conflict of interest was reported by
the author(s).
Funding
This work was in part supported by the Project on Survey
and Discovery of Indigenous Fungal Species of Korea
funded by NIBR, and the Project on Discovery of Fungi
from Freshwater and Collection of Fungarium funded by
NNIBR of the Ministry of Environment (MOE).
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MYCOBIOLOGY 13
